Screening for calcium channel modulators in CLN3 siRNA knock down SH-SY5Y neuroblastoma cells reveals a significant
Kristina An Haack1, Srinivas B Narayan, Haying Li
1Department of Pathology and Laboratory Medicine, Division of Metabolic Disease, The Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA. anhaackk@email.chop.edu
Background:
Defects of the CLN3 gene on chromosome 16p12.1 lead to the juvenile form of neuronal ceroid-lipofuscinosis (JNCL, Batten Disease), the most common recessive inherited neurodegenerative disorder in children. Dysregulation of intracellular calcium homeostasis in the absence of a functional CLN3 protein (CLN3P, Battenin) has been linked to synaptic dysfunction and accelerated apoptosis in vulnerable neuronal cells. Prolonged increase of intracellular calcium concentration is considered to be a significant trigger for neuronal apoptosis and cellular loss in JNCL.
Methods:
We examined the potential effect of 41 different calcium channel modulators on intracellular calcium concentration in CLN3 siRNA knock down SH-SY5Y neuroblastoma cells.
Results:
Six drugs belonging to the group of voltage dependent L-type channel blockers show significant lowering of the increased intracellular calcium levels in CLN3 siRNA knock down cells.
Conclusions:
Our studies provide important new data suggesting possible beneficial effects of the tested drugs on calcium flux regulated pathways in neuronal cell death. Therapeutic intervention in this untreatable disease will likely require drugs that cross the blood-brain barrier as did all of the positively screened drugs in this study.
General Significance:
Better comprehension of the mechanism of neurodegeneration in rare recessive disorders, such as neuronal ceroid-lipofuscinoses, is likely to help to better understand mechanisms involved in more complex genetic neurodegenerative conditions, such as those associated with aging.
Insights
Six drugs that block L-type calcium channels may help treat juvenile neuronal ceroid-lipofuscinosis (JNCL). These drugs reduced high intracellular calcium levels in cells lacking CLN3 protein, offering potential therapeutic avenues for this neurodegenerative disorder.
Area of Science:
- Neuroscience
- Genetics
- Pharmacology
Background:
- Juvenile Neuronal Ceroid Lipofuscinosis (JNCL), also known as Batten Disease, is a severe inherited neurodegenerative disorder in children caused by CLN3 gene defects.
- A key feature of JNCL is the dysregulation of intracellular calcium homeostasis, leading to synaptic dysfunction and neuronal apoptosis.
- Elevated intracellular calcium levels are strongly implicated as a trigger for neuronal cell death in JNCL.
Purpose of the Study:
- To investigate the potential of calcium channel modulators to correct intracellular calcium dysregulation in JNCL.
- To identify specific drug classes that can effectively lower elevated calcium levels in a cellular model of JNCL.
Main Methods:
- Utilized SH-SY5Y neuroblastoma cells with CLN3 gene knockdown via siRNA to mimic JNCL conditions.
- Screened 41 different calcium channel modulators for their effect on intracellular calcium concentration in these cells.
Main Results:
- Identified six drugs, all voltage-dependent L-type calcium channel blockers, that significantly reduced elevated intracellular calcium levels.
- All identified drugs demonstrated the ability to cross the blood-brain barrier, a crucial factor for therapeutic efficacy in neurodegenerative diseases.
Conclusions:
- The study suggests that L-type calcium channel blockers may offer a beneficial therapeutic strategy for JNCL by modulating calcium flux.
- Understanding calcium dysregulation in rare disorders like JNCL can provide insights into mechanisms underlying more common age-related neurodegenerative conditions.
